LOCUS       sequence-75345-e        3027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid compatible with the Golden Gate TALEN cloning system. To 
            clone TAL effectors 7.5 RVDs long..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3027)
  AUTHORS   Boissel S, Jarjour J, Astrakhan A, Adey A, Gouble A, Duchateau P, 
            Shendure J, Stoddard BL, Certo MT, Baker D, Scharenberg AM
  TITLE     megaTALs: a rare-cleaving nuclease architecture for therapeutic 
            genome engineering.
  JOURNAL   Nucleic Acids Res. 2013 Nov 26.
  PUBMED    24285304
REFERENCE   2  (bases 1 to 3027)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2013 Nov 26."
COMMENT     Sequence Label: pFUSC7
FEATURES             Location/Qualifiers
     source          1..3027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             434..1225
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1318..1906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1807..1826
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2060..2077
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2236..2263)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2355..2441)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2495..2872)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
                     HLVRGQVQGETLKTGRRFTTS"
     primer_bind     2505..2521
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2737..2753
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2763..2782
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2763..2781
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2829..2849)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2829..2847)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2868..2884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2868..2884)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2881..2903)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2892..2908
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2916..2922,2923..2940,2941..2946))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2961..2982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg
       61 tggctatcga gacgtctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa
      121 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
      181 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
      241 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
      301 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
      361 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
      421 aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
      481 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
      541 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
      601 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
      661 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
      721 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
      781 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
      841 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
      901 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
      961 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     1021 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     1081 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     1141 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     1201 gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt
     1261 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1321 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1381 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1441 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     1501 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1561 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1621 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1681 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     1741 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1801 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     1861 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     1921 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     1981 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2041 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     2101 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     2161 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     2221 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
     2281 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
     2341 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     2401 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
     2461 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag
     2521 tcttaagcgt ctccccctga acctgacccc ggaccaagtg gtggctatcg agaccggcgc
     2581 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     2641 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     2701 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     2761 cgtaatacga ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcctccagc
     2821 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2881 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3001 cccgctttcc accggtggtc tccgaaa
//
